Abstract: In the current era of combination antiretroviral therapy (cART), there is increased prevalence of HIV-associated neurocognitive disorders (HAND) with about 30-50% of HIV-infected individuals afflicted with varying degrees of neurocognitive impairments, substantially affecting their quality of life. It is well-recognized that despite cART, there is unabated persistence & presence of early HIV protein(s) such as the cytotoxic transactivator of transcription (TAT), that contributes to neuroinflammation. Adding fuel to the mix is the comorbidity of drug abuse in HIV-infected individuals, which further exacerbates microglial activation & thus symptomatology of HAND. Until recently, CNS was considered as an immune-privileged site, however, a recent paradigm shift in our understanding has revealed the role of cytosolic, multiprotein complexes, belonging to the nucleotide-binding domain leucine-rich repeat containing (NLR) protein superfamily, termed as ?inflammasomes,? in various neuroinflammatory diseases. Assembly of these multiprotein complexes activates the proinflammatory caspases (specifically caspase 1), leading, in turn, to the cleavage & release of IL1? & 1L18, consequently resulting in a potent inflammatory response. Among the various NLRs, NLRC5 & NLRP3 are highly expressed in microglia & differentially regulate inflammatory responses. In our efforts to understand the combinatorial effects of HIV Tat & cocaine on Mg activation, we have made several exciting preliminary observations: 1) Exposure of rodent primary Mg to both HIV Tat & cocaine demonstrated significantly downregulated expression of NLRC5, with a concomitant upregulation of NLRP3 and, exacerbated Mg activation compared to cells exposed to either agent alone; 2) Rodent Mg exposed to HIV TAT demonstrated time-dependent upregulation of microRNA (miR)-34a, leading, in turn, to downregulation of its target - NLRC5; 3) Cocaine-mediated activation of Mg involved induction of reactive oxygen species (ROS), that was accompanied with defective mitophagy & subsequent oligomerization of NLRP3 inflammasome complex, leading to induced expression of mature (m)IL1? & IL18; 4) NLRP3 inhibitor - MCC950 mitigated cocaine-mediated activation of Mg. Based on these observations, we hypothesize that the co-operative effects of HIV TAT & cocaine on Mg activation involve two processes: a) downregulated expression of NLRC5 resulting in activation of NF?B (signal 1), leading, in turn, to transcriptional upregulation of NLRP3, pro IL1? & IL18, & b) induction of ROS-mediated defective mitophagy (signal 2), which, in turn, induces activation of NLRP3 inflammasome, resulting in increased cleavage & secretion of mIL1? & IL18. The hypothesis will be tested in two SA: 1) Explore the molecular mechanism(s) underlying HIV & cocaine- mediated induction of the NLRP3 inflammasome leading to Mg activation and 2) To validate in vivo the role of NLRC5/NF?B & ROS/defective mitophagy/NLRP3 axes underlying HIV Tat & cocaine-mediated activation of Mg. Understanding the mechanisms responsible for microglial activation induced by HIV & cocaine will set the stage for the future development of novel therapeutics aimed at dampening the neuroinflammatory responses.